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EFFECTS OF PANAX GINSENG ROOT ON CONDITIONED AVOIDANCE RESPONSE IN RATS
EFFECTS
OF
PANAX
GINSENG
AVOIDANCE
ROOT
RESPONSE
ON IN
Hiroshi SAITO, Moriyoshi TSUCHIYA,
CONDITIONED
RATS Shinichi NAKA
and Keijiro TAKAGI Department of' ChemicalPharmacology, Faculty of' Pharmaceutical Sciences, Unir'crsityof Tokyo, Bunkyo-ku,Tokyo 113, Japan Accepted March 16, 1977
Abstract-Pole climbing and shuttle box tests were employed to study conditioned avoidance response (CAR) and discrimination behaviour in Wistar male rats given extracts from Panax Ginseng root intraperitonealy. Neutral saponins (GNS), a water soluble fraction (GF4) which does not contain saponins, and a lipid soluble fraction (GNo. 5) inhibited CAR and discrimination ability between 500 Hz sound with electric shock (S°) and 1000 Hz sound without shock (Si). Small doses of GNo. 5 and ginsenoside Rg fraction (GRg) produced a slight shortening of the response latency (RL) to the conditioned stimulus in CAR. GNo. 5 produced the incorrect response to SJ. Significant changes in the extinction of CAR were not evident with any fraction. Data from these tests indicate that Panax Ginseng root contains at least three sedative compounds. Panax Ginseng and Japan severity. properties. root
root has been in common
and has been ingested In general
on the central
as a panacea
the compound
Several authors
ly active substances
system (CNS)
of Panax
and shuttle
box tests.
to evaluate
drugs affecting
CAR as a measure between various
and shuttle on motor
of CNS,
Investigators
box tests are employed,
cannot
and muscle
it is necessary
the pole climbing
in rats has been utilized
have also used the inhibition in pharmacological
(5).
to determine
tests were thus adopted
an ataxia and Muscle relaxation.
of
properties curves of
When the pole climbing the effect of given drugs
often indicates
climb the pole or move into another
,Mien ingested,
1)
we employed
tone as escape failure
MATERIALS
data on the com
from the slopes of dose-response
ing rod and suspension produced
of Panax Ginseng
systematic
(CAR)
and differences
can be determined
of the
and sedative
pharmacological
root on the CNS, response
Korea
to differentiate
of CAR and that of the escape behaviour
coordination
when the animal
In an attempt
regardless
stimulant
properties
(1, 2, 3), however,
avoidance
the CNS (4).
of depression
tranquillizers
both the inhibition
Ginseng
Conditioned
to have tonic,
the pharmacological
ponents of Ginseng root were not included.
of years in China,
for all types of disease,
is considered
have reported
nervous
use for thousands
compartment.
a motor
defect
The rotat
to assess the exact dose of drugs which,
AND METHODS
Pole climbing test
Apparatus
The method
described
by Cook and Weidley (4), was used with minor modi
fication.
Test apparatus was a wooden box about 30 x 30 cm square and 60 cm in height,
has a grid floor, a wooden pole and a speaker. rods 0.3 cm in diameter.
The grids are composed of stainless-steel
A forty-five cm long wooden pole with a diameter of 3 cm was
suspended vertically from the top of the box in the center of the square. of the pole did not reach the grid floor.
The bottom end
The speaker attached directly to the top of the
box gives a 500 Hz sound and serves as the conditioned stimulus (CS). The box was in a soundproof enclosure. Training
Male Wistar rats (7-8 weeks old) weighing 150-170 g were used.
At the beginn
ing of the training, the rat was placed in the box for 2 min without CS and electric shock. Then 500 Hz sounds which occurred intermittently once a sec and 0.5 sec in duration, were delivered for 20 sec, and it was confirmed that the rat did not climb the pole.
After this
confirmation, sounds were delivered as CS for 20 sec; sounds only for the first 10 sec and sounds with electric shock (unconditioned stimulus, 0.1 mA, 35 V, AC) for the remaining 10 sec.
Shocks were delivered from the grid at the same time and duration of the sounds.
When the rat climbed the pole, stimuli were immediately terminated.
Rats were exposed to
this situation 60 times a day at an interval of 2 min , for 6 days. Avoidance is defined as climbing during CS only, escape, climbing after electric shock, and escape failure, failure to climb the pole after these stimuli.
The response latency (RL) was considered the time
from presentation of the CS to the response . For each experiment, only rats which showed CAR with a success rate of over 90%; were used. Procedure Experiment 1: during 60 trials.
Trained rats in groups of 10 were administered saline and were tested The results were regarded as control.
During the 3 days after the test,
the rats were given 60 trials at the same time of day, and it was confirmed that the behav ioural baseline did not change.
On the 4th day, the animals were again given 60 trials
soon after the administration of drugs. Experiment 2:
One group was given saline only as a control.
The procedure here was as in experiment 1, except that the number of
trials and the testing time differed.
Trained rats in groups of 10 were given 10 trials twice
daily: 30 and 100 min after the administration of the drugs. Experiment 3:
Trained rats in groups of 6 were exposed to extinction procedure, in
which sounds were delivered without electric shock for 20 sec, 10 trials, 4 tines daily; 10, 40, 70 and 100 min after the injection of extracts from Ginseng root, for 3 days. Experiment 4:
Rats were trained to discriminate between the sounds of 500 Hz (SD)
and those of 1000 Hz (S') which were not followed with electric shock.
The correct re
sponse was defined as the response to climb the pole with SD, or not to climb with S°. Other responses were regarded as incorrect.
Rats in groups of 8 which showed correct response
in rates of over 80% were used. The animals were exposed to 5 trials of SD and those of S' by turns with 20 trials in all, soon after the injection of saline. control.
The results served as
During 3 days after the test, the rats were tested at the same time of day in the
same manner and it was confirmed that the behavioural baseline did not change. 4th day, the animals were tested again soon after the administration of drugs.
On the
2) Shuttle box test Experimental details of the apparatus and training have already been reported (6). Procedure Experiment 5:
Male Wistar rats which could move into another compartment with
only CS (intermittent sounds of 500 Hz) with success rates of over 90 % were used in groups of 10. These animals weighing 180-200 g (8-9 weeks old) were given saline, tested 60 trials at an interval of 1 min and then served as control.
During 3 days after the test, the
rats were tested at the same time of day and it was confirmed that the behaviounral baseline did not change.
On the 4th day, these animals were tested in the same manner soon after
the administration of drugs. Experiment 6:
Rats were trained to discriminate between 500 Hz (SD) and 1000 Hz
(S'). The correct response was defined as the ability to move into another compartment with SD, or not to move with S'. Rats in groups of 6 which showed correct responses at the rate of over 80% were used. The rats were exposed to 5 trials of SD and those of S°, by turns, 20 trials in all at an interval of 1 min, soon after the injection of saline. results were regarded as control.
The
During 3 days after the test, the rats were tested in the
same manner and it was confirmed that the behavioural baseline had not changed.
On
the 4th day, these animals were tested soon after the administration of drugs. 3) Rotating rod test A plastic rod, 10 cm in a diameter, was rotated at a speed of 10 rotations per min. Male Wistar rats, weighing 170-200 g, which could remain on the rotating rod for more than 5 min in two successive trials were used in groups of 6. FIG.
1.
Separation
of Panax
Ginseng
Root.
The test was terminated in
5 min and was repeated 4 times; 30, 60, 90 and 120 min after the injection of drugs.
The
number of rats which fell off the rotating rod within 5 min was counted. 4) Suspension test Male Wistar rats, weighing 160-190 g in groups of 6, were suspended by means of their forepaws to a metallic net of 2 >; 5 cm which was attached to the center of a large vertical glass plate.
Animals which fell from the net within 5 sec were considered to have
a weak muscle tone.
This test was performed 4 times; 30, 60, 90 and 120 min after the
injection of drugs. Drugs tested The preparations
of Panax Ginseng root are shown in Fig. 1.
Four preparations,
namely, neutral saponins (GNS), GNo. 4, ginsenoside Rg (GRg) and GF4, were dissolved in saline.
GRg separated from GNo. 4 contains ginsenoside R-1, R92 and R93. Tile
fraction GF4 is also obtained from the CHC13-eluate of the column chromatography GNo. 4 on silica gel and does not contain saponins. with a drop of tween 80. Methamphetamine
GNo. 5 was suspended in saline
hydrochloride
hydrochloride (CPZ) were used as standard agents.
of
(MA) and chlorpromazine
All drugs were given i.p.
RESULTS 1) Pole climbing test The proportionate
number of rats which showed an escape response was considered
inhibition of CAR and the proportionate number of animals which showed failure to escape as an inhibition of both CAR and escape response. Experiment 1:
Fic. 2.
Effects of the preparations are shown in Fig. 2. GNS in doses of
Effect of Panax Ginseng
Ordinate: Percentage
root on CAR in the pole climbing
test.
The percent of an inhibition of avoidance and escape responses. [---1: of trials of a group on saline which failed to respond to CS but did
respond to US. failed to respond
: Percentage of trials of a group on Panax Ginseng root which to CS but did respond to US. i : Percentage of trials of a
group which failed to respond to US. **: Symbols indicate the significant differ ence of groups between control and drug tested (p<0.01, Student's t-test) and :` : (p<0 .05).
10, 30 and 90 mg/kg, GNo. 4 (100 mg/kg), GF4 (25 and 50 mg/kg) and GNo. 5 (90 mg/kg) induced a significant inhibition of CAR.
At 20 min intervals for 2hr after the injection
of these fractions, the response of each animal was also observed 6 times successively. The percentage of escape response and escape failure or the mean of RL at the time of peak activity within 2 hr allowed for measurements of potency. The maximum effect of GNS was produced about 30 min after the administration. not observed 100 min after the treatment. hr after the treatment.
The inhibitory effect on CAR was
Maximum effect of GNo. 4 occurred within 1
At a close of 50 nmg;kg, GNo. 4, there was a slight shortening of
RL 60-80 min after the treatment (p -,_0.10, Student's t-test). 2 hr.
GF4 inhibited CAR for
GRg (10, 30 and 90 mfg;keg)did not significantly influence CAR or RL, but 10 mg/kg
of GRg slightly reduced RL 40-80 min after the treatment (p<0.10). an inhibition of CAR which lasted for 2 hr.
GNo. 5 produced
Thirty mg/kg of GNo. 5 slightly reduced
the RL 20-60 min after the treatment and such was prolonged 80-120 min after the treat ment (p<0.10). Expcriinent 2:
Results are shown in Fig. 3.
GNS in doses of 10, 30 and 90 nmgjkg
produced an inhibition of CAR and prolonged RL at 30 min after the treatment, but at 100 min after that the effect had disappeared completely. GNo. 4 (50 and 100 nmgkg) had the same effect as GNS on both CAR and RL.
GF_, (12.5, 25 and 50 mg/kg) inhibited
CAR and prolonged RL at 30 min after the treatment, and also at 100 min after that in doses of 25 and 50 mg,'kg.
GRg had no effect on either CAR or RL, though 10 and 30
mg/kg produced a slight reduction of RL 30 min after the treatment (p<0.10).
GNo. 5
(90 mg/kg) inhibited CAR and prolonged RL slignificantly both 30 and 100 min after the treatment. Ten mg/kg of GNo. 5 slightly reduced RL at 100 min after that (p<0.10). Experiment 3:
Influence of GNS in doses of 5 and 10 mg/kg, GNo. 4 (25 and 50
mg/kg), GF4 (5 and 10 m(T;'kg), GRg (10 and 30 iii-;'kg) and GNo. 5 (10 and 30 mg/kg) on the extinction of CAR was also studied.
CPZ in a dose of 2 mg/kg produced a significant
shortening of extinction on the first and second clays (Fig. 4).
Regarding other drugs,
Fin. 3. Effect of Panax Ginseng root on CAR in the pole climbing test. (a): Tests were started 30 min after dosing. (b): Tests were started 100 min after dosing. See Fig. 2 for expression.
FIG. 4.
Effect
of Panax
Ginseng
tinction of response in the test. Ordinate: Extinction avoidance rate or which rats showed
root on ex pole climbing indicates the
FTC. 5. Effect of Panax Ginseng root on dis crimination behaviour in the pole climbing test. Ordinate: Percentage of incorrect .; response. L: Percentage of trials in which rats given saline showed incorrect response. M : Percentage of trials in which rats given drugs showed incorrect response. Symbols as in Fig. 2.
the ratio of trials in CAR to stimuli (Sym
bols as in Fig. 2).
there were no significant changes in the extinction, but 10 mg/kg of GNS and GF4 had a tendency to shorten the extinction in the 3rd or 4th trial on the first day or in the 1st or 2nd trial on the second day.
On the other hand with 10 mg/kg of GRg or GNo. 5 and
2 mg/kg of MA there was a tendency for prolongation of the extinction in the 1st or 2nd trial on the second day. Experiment 4:
Rats were administered the following doses; GNS (5 and 10 mg/kg),
GF4 (5, 10 and 20 mg/kg), GRg (10 and 30 mg/kg) and GNo. 5 (10 and 30 mg/kg). results are shown in Fig. 5.
The
Significant decrease of correct response was observed at the
dose of 10 mg/kg of GNS and GF4, and 30 mg/'kg of GNo. 5. GNS and GF4 inhibited the pole climbing in response to SD like CPZ (3 mg/kg), on the other hand with GNo. 5 and MA (2 mg/kg) the rats climbed in response to S'.
GRg had no effect on discrimination
behaviour. 2)
Shuttle box test Data are presented as shown in the pole climbing test. Experiment 5:
Effects of preparations are shown in Fig. 6.
CPZ in a dose of 3 mg/kg,
GNS (30 mg/ kg), GF4 (50 mg/kg) and GNo. 5 (90 mg/kg) induced a significant inhibition
FIG.
6. in
Effect the
of Panax
shuttle
box
Ginseng
root
on
test
Fig.
2 for
See
CAR ex
FIG.
7.
Effect
of
crimination
pression.
test.
See
Fig.
Panax
Ginseng
behaviour
in
5 for
root the
on
shuttle
dis box
expression.
of CAR and a significant prolongation of RL.
Maximum effect of GNS or GF ,, was pro duced about 30 min after the treatment, and 60 min after that, the effects of these com pounds on CAR and RL had disappeared completely. GRg in doses of 10, 30 and 90 mg/kg had no influence on either CAR or R L, but 10 mgt kg of GRg slightly reduced RL 20-40 min after the treatment (p<0.10).
Inhibitory effect of GNo. 5 (90 mg/kg) on CAR was
evident 60-80 min after the treatment. Experiment 6:
Effects of GNS in doses of 10 and 20 mg/kg , GF4 (10 and 20 ing/kg), GRg (10 and 30 mg/kg) and GNo. 5 (10 and 30 mg/kg) on behaviour regarding discrimina tion between St and SJ were also determined (Fig. 7). Significant decrease of correct re sponse was observed with MA (2 nng/kg), CPZ (3 mg/kg), GNS (20 mg/kg), GF4 (20 mg/kg) and GNo. 5 (30 mg/kg). CPZ, GNS and GF4 inhibited the response to So, and administra tion of MA and GNo. 5 resulted in an incorrect response to S'.
GRg had no effect on
discrimination behaviour. 3)
Rotatin rod acrd .srrsperrsioiltc'sts Effects of drugs an motor coordination and muscle tone were studied in relation to
each of the doses employed in both pole climbing and shuttle box tests . The results in dicated that these fractions had no influence on motor coordination and muscle tone in the doses given herein. DISCUSSION In previous work, we confirmed that GNS Inhibited CAR (7), and GNo. 4 showed a slight CNS-stimulating action (8).
Observation of CAR using pole climbing and shuttle
box tests indicated that GNS produced a significant inhibition of CAR, and a significant decrease of correct response to SD. Large doses of GNo. 4 produced a significant inhibition of CAR, and small doses slightly reduced RL to CS. This inhibition or reduction suggests
that the ingredients activity.
contained
in GNo.
a significant
inhibition
a significant
indicate that CNS-depressing not contain
saponins
CAR or discrimination
Rg,, the main component (9) indicating
nificant inhibition
contains
GNo. 5 produced
substances
reverse pharmacologically
Ginseng
activity.
root
which
activity.
University
activity
It would
extracts
of ginsenoside
fatigued
states in mice
from these results
by GF4 and CNS-stimulating
there was a slight prolongation to S'.
that there
doses
of
Thus GNo. 5 also report (9), GNo.
activities,
and at least two different
activities. to Prof.
S. Shibata,
and Prof. O. Tanaka, of Panax Ginseng
school
Faculty
5
may also possess a
are three different substances
of Pharmaceutical
of Medicine,
Hiroshima
root.
REFERENCES 1) 2) 3) 4) 5) 6) 7) 8) 9)
activity
of RL in small doses and a sig
In our previous
have CNS-depressing
We are grateful
of Tokyo
from
and in smaller
appear
toward reduction
It is considered
a slight reduction
active substances.
which have CNS-stimulating
versity, for providing
in GF4 which does
Administration
increase of incorrect response
to have anti-fatigue
Acknowledgements: Sciences,
of CAR. the recovery
of CAR in large doses, and in addition,
slight CNS-stimulating in Panax
of the extinction
of GRg, accelerated
of CAR and a significant
was demonstrated
and shuttle box
to SD. Such findings
though with GRg there was a tendency
activity of GNo. 4 was represented
of GNo. 4, by GRg.
response
other than GNS are contained
that GRg has CNS-stimulating
that CNS-depressin
of correct
On the other hand, with GRg, there was no change in either
behaviour,
of the RL and a prolongation
GF4 produced
of RL in pole climbing
decrease
components
in itself.
or CNS-stimulating
of GNo. 4 were also examined.
of CAR and prolongation
tests as well as producing
extinction
4 may have CNS-depressing
GF4 and GRg, the components
PETKOV,V.M.: Arzncim.-Forsch. 11, 288 (1961) PETKOV,V.M.: Arzneim.-Forsch. 11, 418 (1961) BREKFIMMAN, 1.1. AND DARDYMOV, I.V.: Proc. Pacific Sci. Congr., 11th, 8, 11 (1966) COOK, L. AND \VEIDLEY,E.F.: Ann. N. 1'. Acad. Sci. 66, 740 (1957) MAEFU,G.: J. Pharm. Pharmacol. 11, 129 (1959) NABATA,H., SArm, I-I. ANDTAKAGI,K.: Japan. J. Pharmacol. 23, 29 (1973) TAKAGI,K. SAITO,1-I.AND NABATA,II.: Japan. J. Plrarmacol. 22, 245 (1972) TAKAGI,K., SAITO, I-1.A.u TS['d inYA, M.: .101)011. .1. I'harmacol. 22, 339 (1972) SAL10,II., YOSIIIDA,Y. ANDTAKAGI,K.: Japan. J. Pharmacol. 24, 119 (1974)
Uni
OF
PANAX
GINSENG
AVOIDANCE
ROOT
RESPONSE
ON IN
Hiroshi SAITO, Moriyoshi TSUCHIYA,
CONDITIONED
RATS Shinichi NAKA
and Keijiro TAKAGI Department of' ChemicalPharmacology, Faculty of' Pharmaceutical Sciences, Unir'crsityof Tokyo, Bunkyo-ku,Tokyo 113, Japan Accepted March 16, 1977
Abstract-Pole climbing and shuttle box tests were employed to study conditioned avoidance response (CAR) and discrimination behaviour in Wistar male rats given extracts from Panax Ginseng root intraperitonealy. Neutral saponins (GNS), a water soluble fraction (GF4) which does not contain saponins, and a lipid soluble fraction (GNo. 5) inhibited CAR and discrimination ability between 500 Hz sound with electric shock (S°) and 1000 Hz sound without shock (Si). Small doses of GNo. 5 and ginsenoside Rg fraction (GRg) produced a slight shortening of the response latency (RL) to the conditioned stimulus in CAR. GNo. 5 produced the incorrect response to SJ. Significant changes in the extinction of CAR were not evident with any fraction. Data from these tests indicate that Panax Ginseng root contains at least three sedative compounds. Panax Ginseng and Japan severity. properties. root
root has been in common
and has been ingested In general
on the central
as a panacea
the compound
Several authors
ly active substances
system (CNS)
of Panax
and shuttle
box tests.
to evaluate
drugs affecting
CAR as a measure between various
and shuttle on motor
of CNS,
Investigators
box tests are employed,
cannot
and muscle
it is necessary
the pole climbing
in rats has been utilized
have also used the inhibition in pharmacological
(5).
to determine
tests were thus adopted
an ataxia and Muscle relaxation.
of
properties curves of
When the pole climbing the effect of given drugs
often indicates
climb the pole or move into another
,Mien ingested,
1)
we employed
tone as escape failure
MATERIALS
data on the com
from the slopes of dose-response
ing rod and suspension produced
of Panax Ginseng
systematic
(CAR)
and differences
can be determined
of the
and sedative
pharmacological
root on the CNS, response
Korea
to differentiate
of CAR and that of the escape behaviour
coordination
when the animal
In an attempt
regardless
stimulant
properties
(1, 2, 3), however,
avoidance
the CNS (4).
of depression
tranquillizers
both the inhibition
Ginseng
Conditioned
to have tonic,
the pharmacological
ponents of Ginseng root were not included.
of years in China,
for all types of disease,
is considered
have reported
nervous
use for thousands
compartment.
a motor
defect
The rotat
to assess the exact dose of drugs which,
AND METHODS
Pole climbing test
Apparatus
The method
described
by Cook and Weidley (4), was used with minor modi
fication.
Test apparatus was a wooden box about 30 x 30 cm square and 60 cm in height,
has a grid floor, a wooden pole and a speaker. rods 0.3 cm in diameter.
The grids are composed of stainless-steel
A forty-five cm long wooden pole with a diameter of 3 cm was
suspended vertically from the top of the box in the center of the square. of the pole did not reach the grid floor.
The bottom end
The speaker attached directly to the top of the
box gives a 500 Hz sound and serves as the conditioned stimulus (CS). The box was in a soundproof enclosure. Training
Male Wistar rats (7-8 weeks old) weighing 150-170 g were used.
At the beginn
ing of the training, the rat was placed in the box for 2 min without CS and electric shock. Then 500 Hz sounds which occurred intermittently once a sec and 0.5 sec in duration, were delivered for 20 sec, and it was confirmed that the rat did not climb the pole.
After this
confirmation, sounds were delivered as CS for 20 sec; sounds only for the first 10 sec and sounds with electric shock (unconditioned stimulus, 0.1 mA, 35 V, AC) for the remaining 10 sec.
Shocks were delivered from the grid at the same time and duration of the sounds.
When the rat climbed the pole, stimuli were immediately terminated.
Rats were exposed to
this situation 60 times a day at an interval of 2 min , for 6 days. Avoidance is defined as climbing during CS only, escape, climbing after electric shock, and escape failure, failure to climb the pole after these stimuli.
The response latency (RL) was considered the time
from presentation of the CS to the response . For each experiment, only rats which showed CAR with a success rate of over 90%; were used. Procedure Experiment 1: during 60 trials.
Trained rats in groups of 10 were administered saline and were tested The results were regarded as control.
During the 3 days after the test,
the rats were given 60 trials at the same time of day, and it was confirmed that the behav ioural baseline did not change.
On the 4th day, the animals were again given 60 trials
soon after the administration of drugs. Experiment 2:
One group was given saline only as a control.
The procedure here was as in experiment 1, except that the number of
trials and the testing time differed.
Trained rats in groups of 10 were given 10 trials twice
daily: 30 and 100 min after the administration of the drugs. Experiment 3:
Trained rats in groups of 6 were exposed to extinction procedure, in
which sounds were delivered without electric shock for 20 sec, 10 trials, 4 tines daily; 10, 40, 70 and 100 min after the injection of extracts from Ginseng root, for 3 days. Experiment 4:
Rats were trained to discriminate between the sounds of 500 Hz (SD)
and those of 1000 Hz (S') which were not followed with electric shock.
The correct re
sponse was defined as the response to climb the pole with SD, or not to climb with S°. Other responses were regarded as incorrect.
Rats in groups of 8 which showed correct response
in rates of over 80% were used. The animals were exposed to 5 trials of SD and those of S' by turns with 20 trials in all, soon after the injection of saline. control.
The results served as
During 3 days after the test, the rats were tested at the same time of day in the
same manner and it was confirmed that the behavioural baseline did not change. 4th day, the animals were tested again soon after the administration of drugs.
On the
2) Shuttle box test Experimental details of the apparatus and training have already been reported (6). Procedure Experiment 5:
Male Wistar rats which could move into another compartment with
only CS (intermittent sounds of 500 Hz) with success rates of over 90 % were used in groups of 10. These animals weighing 180-200 g (8-9 weeks old) were given saline, tested 60 trials at an interval of 1 min and then served as control.
During 3 days after the test, the
rats were tested at the same time of day and it was confirmed that the behaviounral baseline did not change.
On the 4th day, these animals were tested in the same manner soon after
the administration of drugs. Experiment 6:
Rats were trained to discriminate between 500 Hz (SD) and 1000 Hz
(S'). The correct response was defined as the ability to move into another compartment with SD, or not to move with S'. Rats in groups of 6 which showed correct responses at the rate of over 80% were used. The rats were exposed to 5 trials of SD and those of S°, by turns, 20 trials in all at an interval of 1 min, soon after the injection of saline. results were regarded as control.
The
During 3 days after the test, the rats were tested in the
same manner and it was confirmed that the behavioural baseline had not changed.
On
the 4th day, these animals were tested soon after the administration of drugs. 3) Rotating rod test A plastic rod, 10 cm in a diameter, was rotated at a speed of 10 rotations per min. Male Wistar rats, weighing 170-200 g, which could remain on the rotating rod for more than 5 min in two successive trials were used in groups of 6. FIG.
1.
Separation
of Panax
Ginseng
Root.
The test was terminated in
5 min and was repeated 4 times; 30, 60, 90 and 120 min after the injection of drugs.
The
number of rats which fell off the rotating rod within 5 min was counted. 4) Suspension test Male Wistar rats, weighing 160-190 g in groups of 6, were suspended by means of their forepaws to a metallic net of 2 >; 5 cm which was attached to the center of a large vertical glass plate.
Animals which fell from the net within 5 sec were considered to have
a weak muscle tone.
This test was performed 4 times; 30, 60, 90 and 120 min after the
injection of drugs. Drugs tested The preparations
of Panax Ginseng root are shown in Fig. 1.
Four preparations,
namely, neutral saponins (GNS), GNo. 4, ginsenoside Rg (GRg) and GF4, were dissolved in saline.
GRg separated from GNo. 4 contains ginsenoside R-1, R92 and R93. Tile
fraction GF4 is also obtained from the CHC13-eluate of the column chromatography GNo. 4 on silica gel and does not contain saponins. with a drop of tween 80. Methamphetamine
GNo. 5 was suspended in saline
hydrochloride
hydrochloride (CPZ) were used as standard agents.
of
(MA) and chlorpromazine
All drugs were given i.p.
RESULTS 1) Pole climbing test The proportionate
number of rats which showed an escape response was considered
inhibition of CAR and the proportionate number of animals which showed failure to escape as an inhibition of both CAR and escape response. Experiment 1:
Fic. 2.
Effects of the preparations are shown in Fig. 2. GNS in doses of
Effect of Panax Ginseng
Ordinate: Percentage
root on CAR in the pole climbing
test.
The percent of an inhibition of avoidance and escape responses. [---1: of trials of a group on saline which failed to respond to CS but did
respond to US. failed to respond
: Percentage of trials of a group on Panax Ginseng root which to CS but did respond to US. i : Percentage of trials of a
group which failed to respond to US. **: Symbols indicate the significant differ ence of groups between control and drug tested (p<0.01, Student's t-test) and :` : (p<0 .05).
10, 30 and 90 mg/kg, GNo. 4 (100 mg/kg), GF4 (25 and 50 mg/kg) and GNo. 5 (90 mg/kg) induced a significant inhibition of CAR.
At 20 min intervals for 2hr after the injection
of these fractions, the response of each animal was also observed 6 times successively. The percentage of escape response and escape failure or the mean of RL at the time of peak activity within 2 hr allowed for measurements of potency. The maximum effect of GNS was produced about 30 min after the administration. not observed 100 min after the treatment. hr after the treatment.
The inhibitory effect on CAR was
Maximum effect of GNo. 4 occurred within 1
At a close of 50 nmg;kg, GNo. 4, there was a slight shortening of
RL 60-80 min after the treatment (p -,_0.10, Student's t-test). 2 hr.
GF4 inhibited CAR for
GRg (10, 30 and 90 mfg;keg)did not significantly influence CAR or RL, but 10 mg/kg
of GRg slightly reduced RL 40-80 min after the treatment (p<0.10). an inhibition of CAR which lasted for 2 hr.
GNo. 5 produced
Thirty mg/kg of GNo. 5 slightly reduced
the RL 20-60 min after the treatment and such was prolonged 80-120 min after the treat ment (p<0.10). Expcriinent 2:
Results are shown in Fig. 3.
GNS in doses of 10, 30 and 90 nmgjkg
produced an inhibition of CAR and prolonged RL at 30 min after the treatment, but at 100 min after that the effect had disappeared completely. GNo. 4 (50 and 100 nmgkg) had the same effect as GNS on both CAR and RL.
GF_, (12.5, 25 and 50 mg/kg) inhibited
CAR and prolonged RL at 30 min after the treatment, and also at 100 min after that in doses of 25 and 50 mg,'kg.
GRg had no effect on either CAR or RL, though 10 and 30
mg/kg produced a slight reduction of RL 30 min after the treatment (p<0.10).
GNo. 5
(90 mg/kg) inhibited CAR and prolonged RL slignificantly both 30 and 100 min after the treatment. Ten mg/kg of GNo. 5 slightly reduced RL at 100 min after that (p<0.10). Experiment 3:
Influence of GNS in doses of 5 and 10 mg/kg, GNo. 4 (25 and 50
mg/kg), GF4 (5 and 10 m(T;'kg), GRg (10 and 30 iii-;'kg) and GNo. 5 (10 and 30 mg/kg) on the extinction of CAR was also studied.
CPZ in a dose of 2 mg/kg produced a significant
shortening of extinction on the first and second clays (Fig. 4).
Regarding other drugs,
Fin. 3. Effect of Panax Ginseng root on CAR in the pole climbing test. (a): Tests were started 30 min after dosing. (b): Tests were started 100 min after dosing. See Fig. 2 for expression.
FIG. 4.
Effect
of Panax
Ginseng
tinction of response in the test. Ordinate: Extinction avoidance rate or which rats showed
root on ex pole climbing indicates the
FTC. 5. Effect of Panax Ginseng root on dis crimination behaviour in the pole climbing test. Ordinate: Percentage of incorrect .; response. L: Percentage of trials in which rats given saline showed incorrect response. M : Percentage of trials in which rats given drugs showed incorrect response. Symbols as in Fig. 2.
the ratio of trials in CAR to stimuli (Sym
bols as in Fig. 2).
there were no significant changes in the extinction, but 10 mg/kg of GNS and GF4 had a tendency to shorten the extinction in the 3rd or 4th trial on the first day or in the 1st or 2nd trial on the second day.
On the other hand with 10 mg/kg of GRg or GNo. 5 and
2 mg/kg of MA there was a tendency for prolongation of the extinction in the 1st or 2nd trial on the second day. Experiment 4:
Rats were administered the following doses; GNS (5 and 10 mg/kg),
GF4 (5, 10 and 20 mg/kg), GRg (10 and 30 mg/kg) and GNo. 5 (10 and 30 mg/kg). results are shown in Fig. 5.
The
Significant decrease of correct response was observed at the
dose of 10 mg/kg of GNS and GF4, and 30 mg/'kg of GNo. 5. GNS and GF4 inhibited the pole climbing in response to SD like CPZ (3 mg/kg), on the other hand with GNo. 5 and MA (2 mg/kg) the rats climbed in response to S'.
GRg had no effect on discrimination
behaviour. 2)
Shuttle box test Data are presented as shown in the pole climbing test. Experiment 5:
Effects of preparations are shown in Fig. 6.
CPZ in a dose of 3 mg/kg,
GNS (30 mg/ kg), GF4 (50 mg/kg) and GNo. 5 (90 mg/kg) induced a significant inhibition
FIG.
6. in
Effect the
of Panax
shuttle
box
Ginseng
root
on
test
Fig.
2 for
See
CAR ex
FIG.
7.
Effect
of
crimination
pression.
test.
See
Fig.
Panax
Ginseng
behaviour
in
5 for
root the
on
shuttle
dis box
expression.
of CAR and a significant prolongation of RL.
Maximum effect of GNS or GF ,, was pro duced about 30 min after the treatment, and 60 min after that, the effects of these com pounds on CAR and RL had disappeared completely. GRg in doses of 10, 30 and 90 mg/kg had no influence on either CAR or R L, but 10 mgt kg of GRg slightly reduced RL 20-40 min after the treatment (p<0.10).
Inhibitory effect of GNo. 5 (90 mg/kg) on CAR was
evident 60-80 min after the treatment. Experiment 6:
Effects of GNS in doses of 10 and 20 mg/kg , GF4 (10 and 20 ing/kg), GRg (10 and 30 mg/kg) and GNo. 5 (10 and 30 mg/kg) on behaviour regarding discrimina tion between St and SJ were also determined (Fig. 7). Significant decrease of correct re sponse was observed with MA (2 nng/kg), CPZ (3 mg/kg), GNS (20 mg/kg), GF4 (20 mg/kg) and GNo. 5 (30 mg/kg). CPZ, GNS and GF4 inhibited the response to So, and administra tion of MA and GNo. 5 resulted in an incorrect response to S'.
GRg had no effect on
discrimination behaviour. 3)
Rotatin rod acrd .srrsperrsioiltc'sts Effects of drugs an motor coordination and muscle tone were studied in relation to
each of the doses employed in both pole climbing and shuttle box tests . The results in dicated that these fractions had no influence on motor coordination and muscle tone in the doses given herein. DISCUSSION In previous work, we confirmed that GNS Inhibited CAR (7), and GNo. 4 showed a slight CNS-stimulating action (8).
Observation of CAR using pole climbing and shuttle
box tests indicated that GNS produced a significant inhibition of CAR, and a significant decrease of correct response to SD. Large doses of GNo. 4 produced a significant inhibition of CAR, and small doses slightly reduced RL to CS. This inhibition or reduction suggests
that the ingredients activity.
contained
in GNo.
a significant
inhibition
a significant
indicate that CNS-depressing not contain
saponins
CAR or discrimination
Rg,, the main component (9) indicating
nificant inhibition
contains
GNo. 5 produced
substances
reverse pharmacologically
Ginseng
activity.
root
which
activity.
University
activity
It would
extracts
of ginsenoside
fatigued
states in mice
from these results
by GF4 and CNS-stimulating
there was a slight prolongation to S'.
that there
doses
of
Thus GNo. 5 also report (9), GNo.
activities,
and at least two different
activities. to Prof.
S. Shibata,
and Prof. O. Tanaka, of Panax Ginseng
school
Faculty
5
may also possess a
are three different substances
of Pharmaceutical
of Medicine,
Hiroshima
root.
REFERENCES 1) 2) 3) 4) 5) 6) 7) 8) 9)
activity
of RL in small doses and a sig
In our previous
have CNS-depressing
We are grateful
of Tokyo
from
and in smaller
appear
toward reduction
It is considered
a slight reduction
active substances.
which have CNS-stimulating
versity, for providing
in GF4 which does
Administration
increase of incorrect response
to have anti-fatigue
Acknowledgements: Sciences,
of CAR. the recovery
of CAR in large doses, and in addition,
slight CNS-stimulating in Panax
of the extinction
of GRg, accelerated
of CAR and a significant
was demonstrated
and shuttle box
to SD. Such findings
though with GRg there was a tendency
activity of GNo. 4 was represented
of GNo. 4, by GRg.
response
other than GNS are contained
that GRg has CNS-stimulating
that CNS-depressin
of correct
On the other hand, with GRg, there was no change in either
behaviour,
of the RL and a prolongation
GF4 produced
of RL in pole climbing
decrease
components
in itself.
or CNS-stimulating
of GNo. 4 were also examined.
of CAR and prolongation
tests as well as producing
extinction
4 may have CNS-depressing
GF4 and GRg, the components
PETKOV,V.M.: Arzncim.-Forsch. 11, 288 (1961) PETKOV,V.M.: Arzneim.-Forsch. 11, 418 (1961) BREKFIMMAN, 1.1. AND DARDYMOV, I.V.: Proc. Pacific Sci. Congr., 11th, 8, 11 (1966) COOK, L. AND \VEIDLEY,E.F.: Ann. N. 1'. Acad. Sci. 66, 740 (1957) MAEFU,G.: J. Pharm. Pharmacol. 11, 129 (1959) NABATA,H., SArm, I-I. ANDTAKAGI,K.: Japan. J. Pharmacol. 23, 29 (1973) TAKAGI,K. SAITO,1-I.AND NABATA,II.: Japan. J. Plrarmacol. 22, 245 (1972) TAKAGI,K., SAITO, I-1.A.u TS['d inYA, M.: .101)011. .1. I'harmacol. 22, 339 (1972) SAL10,II., YOSIIIDA,Y. ANDTAKAGI,K.: Japan. J. Pharmacol. 24, 119 (1974)
Uni